大鼠海马神经元癫痫样放电模型的构建

目的探讨“无镁细胞外液”诱导体外培养大鼠海马神经元产生癫痫样放电的可行性，以期建立难治性癫痫离体细胞模型。方法选取24h内新生Wistar大鼠，分离海马神经元后进行原代培养，体外培养至第12天时，用“无镁细胞外液”处理3h，应用全细胞膜片钳技术记录海马神经元的放电情况。结果在培养第12天时，神经元突起间彼此接触形成神经网络。在“无镁细胞外液”处理3h后神经元产生稳定的放电，恢复正常细胞培养液培养24h，神经元仍可检测到自发的“癫痫样放电”。结论体外培养第12天海马神经元，在“无钱细胞外液”处理后可形成稳定的自发性癫痫样放电，为今后在细胞分子水平研究癫痫发病机制提供了一种理想模型。

Establishment of the epileptiform discharge model in rat hippocampal neuron

Objective To explore the feasibility of epileptiform discharge of rat hippocampal neurons cultured in vitro and induced by magnesium-free extracellular fluid in order to establish refractory epilepsy model in cultured cells. Methods The neonatal Wistar rats within 24h were used to isolate hippocampal neuron to carry out primary culture in vitro. After 12d, the hippocampal neurons were treated with magnesium free extracellular fluid for 3h, then neuronal discharge activities were recorded by whole cell patch clamping technique. Results After 12 -day cell culture, neuronal dendrites touched each other to form neural network. After 3 - hour treatment by magnesium-free extracellular fluid, the neurons produced stable discharge, however, after 24 - hour treatment with normal cell culture fluid, the spontaneous epileptiform activity in neurons could still be observed. Conclusion The stable spontaneous epileptiform activity can be formed in hippocanlpal neurons cultured in vitro for 12d, after exposed in magnesium free extracellular fluid for 3h, which provides an ideal model for pathogenetic research about epilepsy at cell and molecular level.